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Homozygous lymphoblastoid cell lines representing various Dw subtypes of DR2 were examined for polymorphism at the DQ beta locus by molecular and cellular techniques. The subtypes studied included Dw2, Dw12, and a group heterogenous by cellular typing that we shall refer to as non-Dw2/non-Dw12. Restriction fragment length polymorphism analysis of cell lines representing these subtypes revealed DQ beta-specific patterns consistent with cellular typing. Two-dimensional gel electrophoresis of DQ molecules from representative cell lines revealed a structural polymorphism of DQ beta among the three subtypes. The DQ beta chain migrated to a position that was unique to each subtype and was consistent among various representative cell lines of each subtype. Nucleotide sequence analysis of cDNA clones of DQ beta from Dw2, Dw12, and non-Dw2/non-Dw12 lines confirmed that the variability resided at the genetic level. Variability was found in the form of numerous scattered nucleotide substitutions throughout the first domain of these alleles. The DQ beta gene of the non-Dw2/non-Dw12 cell line AZH was further found to be almost identical with the DQ beta gene of a DR1 line (Bell et al. 1985b), implicating a common evolutionary origin of these alleles. The only difference between these two sequences was due to an apparent gene conversion event at amino acid 57. T-cell cloning experiments resulted in the derivation of Epstein-Barr virus-specific, DQw1-restricted clones that proliferated against only those cell lines that exhibited the DQ beta gene common to AZH and the DR1 cell line. Thus, the polymorphism among DQ beta alleles within DR2 results in subtype-specific restriction.

Type

Journal article

Journal

Immunogenetics

Publication Date

1987

Volume

26

Pages

85 - 91

Keywords

Alleles, Base Sequence, Cell Line, Clone Cells, DNA, Electrophoresis, Polyacrylamide Gel, HLA-D Antigens, HLA-DQ Antigens, HLA-DR Antigens, Lymphocyte Activation, Polymorphism, Restriction Fragment Length, Sequence Homology, Nucleic Acid, T-Lymphocytes